nociceptin  (MedChemExpress)

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: NOP antagonist LY2940094 promotes OPC differentiation . (A) Schematic of the MBP expression-based imaging assay: mouse NPCs from E14.5 embryos were isolated and expanded in the NPC medium to form suspended neurospheres. Two days later, neurospheres were collected and dissociated into single cells and then differentiated to OPC in the OPC medium for 2 days. On day 0, OPC medium was replaced with basal medium (drug to be tested was added in the same day), OPCs were allowed to differentiate in the basal medium for 4 days, OLs were stained with antibody against MBP. (B) Chemical structures of LY2940094. (C, D) The effect of LY2940094 (C) and nociceptin (D) in promoting OPC-to-OL differentiation. The compounds were added into the basal medium in indicated concentrations for 4 days and MBP + cells were counted. (E, F) Statistical analysis (E) and representative images (F) of OPCs induced in the presence of As-2P (100 ​μM), LY2940092 (2.5 ​μM), or the combination of both for 4 days; OLs were stained with antibody against MBP (green). Data are means ​± ​SEM (n ​= ​3), ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001, ∗∗∗∗ p ​< ​0.0001 versus Ctrl group. #### p ​< ​0.0001, LY2940094+As-2P versus LY2940094 only. Scale bars, 100 ​μm.

Article Snippet: LY2940094, JTC-801, SB-612111, BAN ORL24, and nociceptin were purchased from MedChemExpress.

Techniques: Expressing, Imaging, Isolation, Staining

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: NOP antagonist LY2940094 promotes OPC differentiation . (A) Schematic of the MBP expression-based imaging assay: mouse NPCs from E14.5 embryos were isolated and expanded in the NPC medium to form suspended neurospheres. Two days later, neurospheres were collected and dissociated into single cells and then differentiated to OPC in the OPC medium for 2 days. On day 0, OPC medium was replaced with basal medium (drug to be tested was added in the same day), OPCs were allowed to differentiate in the basal medium for 4 days, OLs were stained with antibody against MBP. (B) Chemical structures of LY2940094. (C, D) The effect of LY2940094 (C) and nociceptin (D) in promoting OPC-to-OL differentiation. The compounds were added into the basal medium in indicated concentrations for 4 days and MBP + cells were counted. (E, F) Statistical analysis (E) and representative images (F) of OPCs induced in the presence of As-2P (100 ​μM), LY2940092 (2.5 ​μM), or the combination of both for 4 days; OLs were stained with antibody against MBP (green). Data are means ​± ​SEM (n ​= ​3), ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001, ∗∗∗∗ p ​< ​0.0001 versus Ctrl group. #### p ​< ​0.0001, LY2940094+As-2P versus LY2940094 only. Scale bars, 100 ​μm.

Article Snippet: LY2940094, recently renamed as BTRX-246040, was originally developed by Eli Lilly as an orally available specific NOPR antagonist with high affinity and antagonist potency [ , ].

Techniques: Expressing, Imaging, Isolation, Staining

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: LY2940094 promotes gradual expression of OL lineage markers and OL maturation . (A–B) Time-dependent effect of LY2940094 (2.5 ​μM) in OPCs differentiation. OPCs were treated with LY2940094 for the indicated time period, MBP + cells (green) were counted (A), or the MBP levels in the cells were analyzed with Western blot (B). (C) Representative images of cells expressing O4 (green) during OPC-to-OL differentiation in the presence of LY2940094 (2.5 ​μM). (D) Statistical analysis of O4 + cells according to their morphologies which are defined as simple, complex or membrane structure. (E and F) Representative images (E) and statistical analysis (F) of MBP + cells during OPC-to-OL differentiation in the presence of LY2940094 (2.5 ​μM). Data are means ​± ​SEM (n ​= ​3), ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001 versus Ctrl. Scale bars, 100 ​μm.

Article Snippet: LY2940094, recently renamed as BTRX-246040, was originally developed by Eli Lilly as an orally available specific NOPR antagonist with high affinity and antagonist potency [ , ].

Techniques: Expressing, Western Blot, Membrane

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: LY2940094 boosts in vitro myelination . (A-C) The effect of LY2940094 in promoting the differentiation of primary NG2 + OPCs isolated from the postnatal mice with Anti-AN2 MicroBeads. Statistical analysis (A) and representative images (B) of MBP + cells (green) and Western blot analysis of MBP expression (C) after differentiation for 3 days. Scale bars, 100 ​μm. (D) Representative images of in vitro myelination with co-culture of DRG neurons and primary OPCs for 6 days in the presence of LY2940094 (10 ​μM). The cells were immunostained for neurofilament NF-200 (red) and MBP (green). Arrows indicate myelinated axons (double positive for NF-200 and MBP). Scale bars, 50 ​μm. (E) Statistical analysis of myelinated axons in (D). Data are means ​± ​SEM (n ​= ​3), ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001, ∗∗∗∗ p ​< ​0.0001 versus Ctrl group.

Article Snippet: LY2940094, recently renamed as BTRX-246040, was originally developed by Eli Lilly as an orally available specific NOPR antagonist with high affinity and antagonist potency [ , ].

Techniques: In Vitro, Isolation, Western Blot, Expressing, Co-Culture Assay

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: LY2940094 promotes myelin repair in cuprizone induced demyelination mouse model . (A) Schematic of the cuprizone induced demyelination/remyelination mouse model. C57BL/6 mice were fed with a diet containing 0.2% cuprizone for 5 weeks to induce demyelination. Following cuprizone withdrawal, the mice were orally treated with vehicle or LY2940094 (30 ​mg/kg) for 1, 2, or 3 weeks (wks). (B) Representative images of the corpus callosum region stained with Luxol Fast Blue at indicated timepoints. Scale bars, 200 ​μm. (C) Statistical analysis of the demyelinating areas in (Data are means ​± ​SEM (n ​= ​5), ∗ p ​< ​0.05, versus Ctrl group, 5 mice per group, five sections of the corpus callosum region from each mouse were analyzed (Student's t -test)). Representative images of MBP (D) and GST-π (Ε) staining of the corpus callosum region of mice treated with control (Ctrl) or LY2940094 (30 ​mg/kg) for 2 weeks. (F) Statistical analysis of MBP fluorescent intensity and GST-π positive cell number in D and E (Data are means ​± ​SEM (n ​= ​5), ∗ p ​< ​0.05, ∗∗∗ p ​< ​0.001 versus Ctrl group, 5 mice per group, 3–4 sections of the corpus callosum region from each mouse were analyzed (Student's t -test)). Scale bars, 200 ​μm.

Article Snippet: LY2940094, recently renamed as BTRX-246040, was originally developed by Eli Lilly as an orally available specific NOPR antagonist with high affinity and antagonist potency [ , ].

Techniques: Staining, Control

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: LY2940094 promotes oligodendrocytes generation by an NOPR independent pathway . (A-C) Effect of NOPR antagonists JTC-804 (A), SB-612111(B) and BAN ORL24 (C) in OPC-to-OL differentiation. NPC-derived OPCs were differentiated in the presence of various concentration of these compounds for 4 days, OLs were stained with antibody against MBP (red) and percentage of MBP + cells were analyzed. (D–F) Effect of NOPR knockdown in OPC-to-OL differentiation. NPC-derived OPCs were infected with indicated lentivirus or lentivirus mixture carrying shRNAs that target Oprl1 , then allow to differentiate in the presence of solvent control, LY2940094 (2.5 ​μM) and As-2P (100 ​μM) for 4 days, and MBP + cells were pictured (D) and analyzed (F), knockdown efficiency was quantified with qPCR analysis (E). (G) NPC-derived OPCs were treated with LY2940094 (2.5 ​μM) for various time, cell lysates were immunoblotted with the indicated antibodies. PD (ERK1/2 inhibitor PD0325901) was used as positive control. (H, I) Quantitative real time PCR analysis of the expression of myelin-related genes (H) and OLs differentiation-associated transcription factors (I) in LY2940094 treated or control cells. Results were normalized to β-actin and then normalized to the control (Ctrl) group. Data are means ​± ​SEM (n ​= ​3), ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001, ∗∗∗∗ p ​< ​0.0001 versus Ctrl group.

Article Snippet: LY2940094, recently renamed as BTRX-246040, was originally developed by Eli Lilly as an orally available specific NOPR antagonist with high affinity and antagonist potency [ , ].

Techniques: Derivative Assay, Concentration Assay, Staining, Knockdown, Infection, Solvent, Control, Positive Control, Real-time Polymerase Chain Reaction, Expressing

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: NOP antagonist LY2940094 promotes OPC differentiation . (A) Schematic of the MBP expression-based imaging assay: mouse NPCs from E14.5 embryos were isolated and expanded in the NPC medium to form suspended neurospheres. Two days later, neurospheres were collected and dissociated into single cells and then differentiated to OPC in the OPC medium for 2 days. On day 0, OPC medium was replaced with basal medium (drug to be tested was added in the same day), OPCs were allowed to differentiate in the basal medium for 4 days, OLs were stained with antibody against MBP. (B) Chemical structures of LY2940094. (C, D) The effect of LY2940094 (C) and nociceptin (D) in promoting OPC-to-OL differentiation. The compounds were added into the basal medium in indicated concentrations for 4 days and MBP + cells were counted. (E, F) Statistical analysis (E) and representative images (F) of OPCs induced in the presence of As-2P (100 ​μM), LY2940092 (2.5 ​μM), or the combination of both for 4 days; OLs were stained with antibody against MBP (green). Data are means ​± ​SEM (n ​= ​3), ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001, ∗∗∗∗ p ​< ​0.0001 versus Ctrl group. #### p ​< ​0.0001, LY2940094+As-2P versus LY2940094 only. Scale bars, 100 ​μm.

Article Snippet: LY2940094, JTC-801, SB-612111, BAN ORL24, and nociceptin were purchased from MedChemExpress.

Techniques: Expressing, Imaging, Isolation, Staining

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: LY2940094 promotes gradual expression of OL lineage markers and OL maturation . (A–B) Time-dependent effect of LY2940094 (2.5 ​μM) in OPCs differentiation. OPCs were treated with LY2940094 for the indicated time period, MBP + cells (green) were counted (A), or the MBP levels in the cells were analyzed with Western blot (B). (C) Representative images of cells expressing O4 (green) during OPC-to-OL differentiation in the presence of LY2940094 (2.5 ​μM). (D) Statistical analysis of O4 + cells according to their morphologies which are defined as simple, complex or membrane structure. (E and F) Representative images (E) and statistical analysis (F) of MBP + cells during OPC-to-OL differentiation in the presence of LY2940094 (2.5 ​μM). Data are means ​± ​SEM (n ​= ​3), ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001 versus Ctrl. Scale bars, 100 ​μm.

Article Snippet: LY2940094, JTC-801, SB-612111, BAN ORL24, and nociceptin were purchased from MedChemExpress.

Techniques: Expressing, Western Blot, Membrane

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: LY2940094 boosts in vitro myelination . (A-C) The effect of LY2940094 in promoting the differentiation of primary NG2 + OPCs isolated from the postnatal mice with Anti-AN2 MicroBeads. Statistical analysis (A) and representative images (B) of MBP + cells (green) and Western blot analysis of MBP expression (C) after differentiation for 3 days. Scale bars, 100 ​μm. (D) Representative images of in vitro myelination with co-culture of DRG neurons and primary OPCs for 6 days in the presence of LY2940094 (10 ​μM). The cells were immunostained for neurofilament NF-200 (red) and MBP (green). Arrows indicate myelinated axons (double positive for NF-200 and MBP). Scale bars, 50 ​μm. (E) Statistical analysis of myelinated axons in (D). Data are means ​± ​SEM (n ​= ​3), ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001, ∗∗∗∗ p ​< ​0.0001 versus Ctrl group.

Article Snippet: LY2940094, JTC-801, SB-612111, BAN ORL24, and nociceptin were purchased from MedChemExpress.

Techniques: In Vitro, Isolation, Western Blot, Expressing, Co-Culture Assay

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: LY2940094 promotes myelin repair in cuprizone induced demyelination mouse model . (A) Schematic of the cuprizone induced demyelination/remyelination mouse model. C57BL/6 mice were fed with a diet containing 0.2% cuprizone for 5 weeks to induce demyelination. Following cuprizone withdrawal, the mice were orally treated with vehicle or LY2940094 (30 ​mg/kg) for 1, 2, or 3 weeks (wks). (B) Representative images of the corpus callosum region stained with Luxol Fast Blue at indicated timepoints. Scale bars, 200 ​μm. (C) Statistical analysis of the demyelinating areas in (Data are means ​± ​SEM (n ​= ​5), ∗ p ​< ​0.05, versus Ctrl group, 5 mice per group, five sections of the corpus callosum region from each mouse were analyzed (Student's t -test)). Representative images of MBP (D) and GST-π (Ε) staining of the corpus callosum region of mice treated with control (Ctrl) or LY2940094 (30 ​mg/kg) for 2 weeks. (F) Statistical analysis of MBP fluorescent intensity and GST-π positive cell number in D and E (Data are means ​± ​SEM (n ​= ​5), ∗ p ​< ​0.05, ∗∗∗ p ​< ​0.001 versus Ctrl group, 5 mice per group, 3–4 sections of the corpus callosum region from each mouse were analyzed (Student's t -test)). Scale bars, 200 ​μm.

Article Snippet: LY2940094, JTC-801, SB-612111, BAN ORL24, and nociceptin were purchased from MedChemExpress.

Techniques: Staining, Control

Journal: Neurotherapeutics

Article Title: LY2940094, an NOPR antagonist, promotes oligodendrocyte generation and myelin recovery in an NOPR independent manner

doi: 10.1016/j.neurot.2024.e00424

Figure Lengend Snippet: LY2940094 promotes oligodendrocytes generation by an NOPR independent pathway . (A-C) Effect of NOPR antagonists JTC-804 (A), SB-612111(B) and BAN ORL24 (C) in OPC-to-OL differentiation. NPC-derived OPCs were differentiated in the presence of various concentration of these compounds for 4 days, OLs were stained with antibody against MBP (red) and percentage of MBP + cells were analyzed. (D–F) Effect of NOPR knockdown in OPC-to-OL differentiation. NPC-derived OPCs were infected with indicated lentivirus or lentivirus mixture carrying shRNAs that target Oprl1 , then allow to differentiate in the presence of solvent control, LY2940094 (2.5 ​μM) and As-2P (100 ​μM) for 4 days, and MBP + cells were pictured (D) and analyzed (F), knockdown efficiency was quantified with qPCR analysis (E). (G) NPC-derived OPCs were treated with LY2940094 (2.5 ​μM) for various time, cell lysates were immunoblotted with the indicated antibodies. PD (ERK1/2 inhibitor PD0325901) was used as positive control. (H, I) Quantitative real time PCR analysis of the expression of myelin-related genes (H) and OLs differentiation-associated transcription factors (I) in LY2940094 treated or control cells. Results were normalized to β-actin and then normalized to the control (Ctrl) group. Data are means ​± ​SEM (n ​= ​3), ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p ​< ​0.001, ∗∗∗∗ p ​< ​0.0001 versus Ctrl group.

Article Snippet: LY2940094, JTC-801, SB-612111, BAN ORL24, and nociceptin were purchased from MedChemExpress.

Techniques: Derivative Assay, Concentration Assay, Staining, Knockdown, Infection, Solvent, Control, Positive Control, Real-time Polymerase Chain Reaction, Expressing